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TWISTDX RPA試劑盒（（加MMLV和抑制劑）操作建議
閱讀：1853 發布時間：2020-5-12
英國TwistDx公司（前身為ASM科技有限公司）成立于1999年，基于英國劍橋Babraham研究院建立的生物技術公司。該公司通過突破等溫DNA擴增，開發的重組酶聚合酶擴增*技術（RPA），被譽為DNA診斷領域的革命性創新。
RT試劑盒舉例
（1）Revert Reverse Transcriptase(200U/uL)(Thermo EP0441)
（2）RNA Inhibitor (40,000 U/mL)( Enzymatics Y9240L
TWISTDX廠家回復
“We recommend using similar RT amounts to that of a PCR reaction of the same volume, according to your chosen manufacturer’s instructions.
We also suggest that you run reactions at 40°C and delay agitation of the reaction by a minute to give the RT time to work, otherwise you just run the reaction as you would a normal TwistAmp® reaction (it’s a one-step process).
It’s also advisable to add RNase Inhibitor to any reaction where RNA is the target material. Any commercially available RNase Inhibitor will be suitable.
If the M-MLV RT that you are using comes with a separate dilution/reaction buffer, we suggest not using it as it will lower the concentration of the enzyme.
The “delay agitation of the reaction by a minute” refers to the first mixing step per the protocol in the manual (Step 4). So wait 5 minutes instead of 4 minutes before “vortex, spin down briefly, and return the samples to the reader ensuring that the tubes are returned to their original positions in the block.”
If we were to try and use it again, we would do a titration to determine optimal quantity to add in the new formulation reactions (we haven’t used it since we changed the formulation of the reactions a couple of years ago).
When using RNase Inhibitor. We have used 1µg /50µl reaction, some R&D users have used 2µg /50ul, which didn’t appear to have a negative effect on the performance in our own assay.
For this : RNAse Inhibitor, that manufacture uses 10 ul of the RNAse solution in 50 ul reaction volumes for activity determination, but your customer can try different amounts 5ul or 20ul to see what will give the best results. For this product, references of 50 ul PCR reaction volumes would also be reasonable.
TwistDx以此技術為基礎生產的核酸擴增類產品，能在恒定且較低的溫度下，僅用10~20分鐘，進行單分子核酸檢測。這一技術對環境、硬件設施的要求很低，對于生物防護、水體檢驗、食品檢驗、醫療診斷、微流體、獸醫等方面都有良好的應用。
RPA擴增的各種體系試劑盒中，含有DNA、RNA擴增所需的所有試劑，研究者只要準備好引物和模板就行了。擴增結果可以分別通過凝膠電泳檢測、熒光定量檢測或試紙條檢測，產物純化后可用于下游研究。
默瑞生物
默瑞(上海)生物科技有限公司，作為專業的生命科學產品推廣與服務商，以“創新，專業，共贏”的理念，不斷強化服務體系，提升客戶體驗，以莫納生物產品為主線，結合國內外優質產品提供高標準完整解決方案，幫助科研客戶和生物技術企業高效創新，加速轉化，創造更多優質產品，提升社會福祉，實現多方共贏。
默瑞生物創立以來，已經陸續導入TwistDx,enzymatics,KAPABIOSYSTEMS,BioDynami,Jackson,Echelon,ChromoTek,Nanocs,Biotechrabbit,LeeBiosolutions, magtivio等優質品牌，逐步培育成為專業用戶的選擇。